Abstract
Delivery of proteins has been regarded as the safest and most useful application in therapeutic application of stem cells, because proteins can regulate gene expression transiently without any genomic alteration. However, it is difficult to accurately measure efficiency or quantity of intracellular protein uptake. Here, we performed a comparison study of cell-penetrating peptide (CPP)-conjugated protein delivery system using seven arginine and Streptolysin O (SLO)-mediated system. To compare CPP- and SLO-mediated protein delivery systems, we used GFP and ESRRB protein, which is known to regulate pluripotency-related genes, for delivery into human bone marrow stromal cells (hBMSCs) and human testicular stromal cells (hTSCs). We found that CPP-conjugated protein delivery was more efficient, lower cytotoxicity, and higher biological activity than SLO-mediated protein delivery system. These results suggest that delivery of CPP-conjugated proteins is an efficient tool for introducing biologically active proteins into cells and may have important implications in clinical cell-based therapy.